Pyrimidine derivatives possessing cell-cycle inhibitors activity

ABSTRACT

Compounds of the formula (I), and pharmaceutically acceptable salts and in vivo hydrolysable esters thereof are described. Also described are processes for their preparation and their use as medicaments, particularly medicaments for producing a cell cycle inhibitory (anti cell proliferation) effect in a warm blooded animal, such as man.

This application is a 371 of PCT/GB04/02019 filed May 12, 2004.

The invention relates to pyrimidine derivatives, or pharmaceuticallyacceptable salts or in vivo hydrolysable esters thereof, which possesscell-cycle inhibitory activity and are accordingly useful for theiranti-cell-proliferation (such as anti-cancer) activity and are thereforeuseful in methods of treatment of the human or animal body. Theinvention also relates to processes for the manufacture of saidpyrimidine derivatives, to pharmaceutical compositions containing themand to their use in the manufacture of medicaments of use in theproduction of an anti-cell-proliferation effect in a warm-blooded animalsuch as man.

The cell cycle is fundamental to the survival, regulation andproliferation of cells and is highly regulated to ensure that each stepprogresses in a timely and orderly manner. The progression of cellsthrough the cell cycle arises from the sequential activation andde-activation of several members of the cyclin-dependent kinase (CDK)family. The activation of CDKs is dependent on their interaction with afamily of intracellular proteins called cyclins Cyclins bind to CDKs andthis association is essential for CDK (such as CDK1, CDK2, CDK4 and/orCDK6) activity within the cell. Different cyclins are expressed anddegraded at different points in the cell cycle to ensure that activationand inactivation of CDKs occurs in the correct order for progressionthrough the cell cycle.

Moreover, CDKs appear to be downstream of a number of oncogenesignalling pathways. Disregulation of CDK activity by upregulation ofcyclins and/or deletion of endogenous inhibitors appears to be animportant axis between mitogenic signalling pathways and proliferationof tumour cells.

Accordingly it has been recognised that an inhibitor of cell cyclekinases, particularly inhibitors of CDK1, CDK2 and/or CDK4 (whichoperate at the G2/M, G1-S-G2/M and G1-S phase respectively) should be ofvalue as a selective inhibitor of cell proliferation, such as growth ofmammalian cancer cells.

WO 02/20512, WO 03/076435, WO 03/076436, WO 03/076434 and WO 03/076433describe certain 2-anilino-4-imidazolylpyrimidine derivatives thatinhibit the effect of cell cycle kinases. The present invention is basedon the discovery that a novel group of 2-anilino-4-bicyclicpyrimidinessurprisingly inhibit the effects of cell cycle kinases showingselectivity for CDK1, CDK2and CDK3, and thus possessanti-cell-proliferation properties. The compounds of the presentinvention are not specifically disclosed in any of the aboveapplications. Such properties are expected to be of value in thetreatment of disease states associated with aberrant cell cycles andcell proliferation such as cancers (solid tumours and leukerias),fibroproliferative and differentiative disorders, psoriasis, rheumatoidarthritis, Kaposi's sarcoma, haemangioma, acute and chronicnephropathies, atheroma, atherosclerosis, arterial restenosis,autoimmune diseases, acute and chronic inflammation, bone diseases andocular diseases with retinal vessel proliferation.

Accordingly, the present invention provides a compound of formula (I):

wherein:

Ring A is carbocyclyl or heterocyclyl fused to the imidazole ring;

R¹ is halo, nitro, cyano, hydroxy, amino, carboxy, carbamoyl, mercapto,C₁₋₆alkyl, C₁₋₆alkoxy, C₂₋₆alkenyl or C₂₋₆alkynyl;

p is 0-4; wherein the values of R¹ may be the same or different;

R² is sulphamoyl or a group R^(a)—R^(b)—;

q is 0-2; wherein the values of R² maybe the same or different; andwherein p+q=0-5;

R³ is halo, nitro, cyano, hydroxy, trifluoromethyl, trifluoromethoxy,amino, carboxy, carbamoyl, mercapto, sulphamoyl, C₁₋₃alkyl, C₂₋₃alkenyl,C₂₋₃alkynyl, C₁₋₃alkoxy, C₁₋₃alkanoyl, N—(C₁₋₃alkyl)amino,N)N—(C₁₋₃alkyl)₂amino, C₁₋₃alkanoylamino, N—(C₁₋₃alkyl)carbamoyl,N,N—(C₁₋₃alkyl)₂carbamoyl, C₁₋₃alkylS(O)_(a) wherein a is 0 to 2,N—(C₁₋₃alkyl)sulphamoyl or N,N—(C₁₋₃alkyl)₂sulphamoyl; wherein R³ may beoptionally substituted on carbon by one or more R^(c);

n is 0 to 2, wherein the values of R³ may be the same or different;

R⁴ and R⁵ are independently selected from hydrogen, halo, nitro, cyano,hydroxy, trifluoromethoxy, amino, carboxy, carbamoyl, mercapto,sulphamoyl, C₁₋₆alkyl, C₂₋₆alkenyl, C₂alkynyl, C₁₋₆alkoxy, C₁₋₆alkanoyl,C₁₋₆alkanoyloxy, N—(C₁₋₆yl)amino, N,N—(C₁₋₆alkyl)₂amino,C₁₋₆alkanoylamino, N—(C₁₋₆alkyl)carbamoyl, N,N—C₁₋₆alkyl)₂carbamoyl,C₁₋₆alkylS(O)_(a) wherein a is 0 to 2, C₁₋₆alkoxycarbonyl,N—(C₁₋₆alkyl)sulphamoyl, N,N—(C₁₋₆alkyl)₂sulphamoyl,C₁₋₆alkylsulphonylamino, C₃₋₈cycloalkyl or a 4-7 membered saturatedheterocyclic group; wherein R⁴ and R⁵ may be optionally substituted oncarbon by one or more R^(e); and wherein if said 4-7 membered saturatedheterocyclic group contains an —NH— moiety that nitrogen may beoptionally substituted by a group selected from R^(f);

m is 0-4; wherein the values of R⁴ may be the same or different;

R^(a) is selected from C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₃₋₈cycloalkyl, C₃₋₈cycloalkylC₁₋₆allyl, phenyl, a heterocyclic group,phenylC₁₋₆alkyl or (heterocyclic group)C₁₋₆alkyl; wherein R^(a) may beoptionally substituted on carbon by one or more R^(g); and wherein ifsaid heterocyclic group contains an —NH— moiety that nitrogen may beoptionally substituted by a group selected from R^(h);

R^(b) is —C(O)—, —N(R^(m))C(O)—, —C(O)N(R^(m))—, —S(O)_(r)—,—OC(O)N(R^(m))SO₂—, —SO₂N(R^(m))— or —N(R^(m))SO₂—; wherein R^(m) ishydrogen or C₁₋₆alkyl optionally substituted by one or more R^(j) and ris 1-2;

R^(g) and R^(i) are independently selected from halo, nitro, cyano,hydroxy, amino, carboxy, carbamoyl, mercapto, sulphamoyl, C₁₋₆alkyl,C₂₋₆alkenyl, C₂₋₆alkynyl, C₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkoxy,C₁₋₆alkoxyC₁₋₆alkoxyC₁₋₆alkoxy, C₁₋₆alkanoyl, C₁₋₆alkanoyloxy,N—(C₁₋₆alkyl)amino, N,N—(C₁₋₆alkyl)₂amino, C₁₋₆alkanoylamino,N-(C₁₋₆alkyl)carbamoyl, N,N—(C₁₋₆alkyl)₂carbamoyl, C₁₋₆alkylS(O)_(a)wherein a is 0 to 2, C₁₋₆alkoxycarbonyl, N—(C₁₋₆alkyl)sulphamoyl,N,N—(C₁₋₆alkyl)₂sulphamoyl, C₁₋₆alkylsulphonylamino, C₃₋₈cycloalkyl,phenyl, heterocyclic group, phenylC₁₋₆alkyl-R^(o)—, (heterocyclicgroup)C₁₋₆alkyl-R^(o)—, phenyl-R^(o)— or (heterocyclic group)-R^(o)—;wherein R^(g) and R^(i) independently of each other may be optionallysubstituted on carbon by one or more R^(j); and wherein if saidheterocyclic group contains an —NH— moiety that nitrogen may beoptionally substituted by a group selected from R^(k);

R^(o) is —O—, —N(R^(P))—, —C(O)—, —N(R^(P))C(O)—, —C(O)N(R^(P))—,—S(O)_(s)—, —SO₂N(R^(P))— or —N(R^(P))SO₂—; wherein R^(P) is hydrogen orC₁₋₆alkyl and s is 0-2;

R^(f), R^(h) and R^(k) are independently selected from C₁₋₄alkyl,C₁₋₄alkanoyl, C₁₋₄alkylsulphonyl, C₁₋₄alkoxycarbonyl, carbamoyl,N—(C₁₋₄alkyl)carbamoyl, N,N—(C₁₋₄alkyl)carbamoyl, benzyl,benzyloxycarbonyl, benzoyl and phenylsulphonyl; wherein R^(f), R^(h) andR^(k) independently of each other may be optionally substituted oncarbon by on or more R^(l); and

R^(c), R^(e), R^(l) and R^(j) are independently selected from halo,nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino,carboxy, carbamoyl, mercapto, sulphamoyl, methyl, ethyl, methoxy,ethoxy, acetyl acetoxy, methylamino, ethylamino, dimethylamino,diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl,N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl,N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulphinyl,ethylsulphinyl, mesyl, ethylsulphonyl, methoxycarbonyl, ethoxycarbonyl,N-methylsulphamoyl, N-ethylsulphamoyl, N,N-dimethylsulphamoyl,N,N-diethylsulphamoyl or N-methyl-N-ethylsulphamoyl;

or a pharmaceutically acceptable salt or an in vivo hydrolysable esterthereof.

In this specification the term “alkyl” includes both straight andbranched chain alkyl groups but references to individual alkyl groupssuch as “propyl” are specific for the straight chain version only. Forexample, “C₁₋₆alkyl” includes C₁₋₄alkyl, C₁₋₃alkyl, propyl, isopropyland t-butyl. However, references to individual alkyl groups such as‘propyl’ are specific for the straight chained version only andreferences to individual branched chain alkyl groups such as ‘isopropyl’are specific for the branched chain version only. A similar conventionapplies to other radicals, for example “phenylC₁₋₆alkyl” includesphenylC₁₋₄alkyl, benzyl, 1-phenylethyl and 2-phenylethyl. The term“halo” refers to fluoro, chloro, bromo and iodo.

Where optional substituents are chosen from “one or more” groups it isto be understood that this definition includes all substituents beingchosen from one of the specified groups or the substituents being chosenfrom two or more of the specified groups.

A “carbocyclyl” is a saturated 5-7 membered carbon ring fused to theimidazole ring of formula (I) as shown; wherein a —CH₂— group canoptionally be replaced by a —C(O)—. Suitable values of “carbocyclyl” arecyclopentyl, cyclohexyl and cycloheptyl forming6,7-dihydro-5H-pyrrolo[1,2-a]imidazole;5,6,7,8-tetrahydroimidazo[1,2-a]pyridine; and6,7,8,9-tetrahydro-5H-imidazo[1,2-a]azepine ring systems respectivelywhen fused to the imidazole ring of formula (I).

A “heterocyclyl” is a saturated 5-7 membered ring of which at least oneatom is chosen from nitrogen, sulphur or oxygen, which is fused to theimidazole ring of formula (I) as shown; wherein a —CH₂— group canoptionally be replaced by a —C(O)—. Suitable values of “heterocyclyl”are pyrrolidinyl, tetrahydropyranyl, piperidinyl and homopiperidinylforming for example 2,3-dihydro-1H-imidazo[1,2-a]imidazole,5,6-dihydro-8H-imidazo[2,1-c][1,4]oxazine;5,6,7,8-tetrahydroimidazo[1,2-a]pyrazine; and6,7,8,9-tetrahydro-5H-imidazo[1,2-a][1,4]diazepine ring systemsrespectively when fused to the imidazole ring of formula (I).

A “heterocyclic group” is a saturated, partially saturated orunsaturated, mono or bicyclic ring containing 4-12 atoms of which atleast one atom is chosen from nitrogen, sulphur or oxygen, which may,unless otherwise specified, be carbon or nitrogen linked, wherein a—CH₂— group can optionally be replaced by a —C(O)—, a ring nitrogen atommay optionally bear a C₁₋₆alkyl group and form a quaternary compound ora ring nitrogen and/or sulphur atom may be optionally oxidised to formthe N-oxide and or the S-oxides. Examples and suitable values of theterm “heterocyclic group” are morpholino, piperidyl, pyridyl, pyranyl,pyrrolyl, isothiazolyl, indolyl, quinolyl, thienyl, 1,3-benzodioxolyl,thiadiazolyl, piperazinyl, thiazolidinyl, pyrrolidinyl, thiomorpholino,pyrrolinyl, homopiperazinyl, 3,5-dioxapiperidinyl, tetrahydropyranyl,imidazolyl, pyrimidyl, pyrazinyl, pyridazinyl, isoxazolyl,N-methylpyrrolyl, 4-pyridone, 1-isoquinolone, 2-pyrrolidone,4-thiazolidone, pyridine-N-oxide and quinoline-N-oxide. Preferably a“heterocyclic group” is a saturated, partially saturated or unsaturated,mono or bicyclic ring containing 5 or 6 atoms of which at least one atomis chosen from nitrogen, sulphur or oxygen, it may, unless otherwisespecified, be carbon or nitrogen linked, a —CH₂— group can optionally bereplaced by a —C(O)— and a ring sulphur atom may be optionally oxidisedto form the S-oxides. More preferably a “heterocyclic group” istetrahydrofiryl, pyridyl, pyrrolidinonyl, morpholino, imidazolyl,piperidinyl or pyrrolidinyl. Particularly a “heterocyclic group” istetrahydrofiryl or morpholino. In another aspect of the invention,particularly a “heterocyclic group” is tetrahydrofuran-2-yl,2-oxopyrrolidin-1-yl, furan-2-yl, oxazolyl, morpholino, piperidinyl,thiazolyl, pyrazinyl, isoxazolyl, tetrahydropyran, pyridyl, isoxazolyl,isothiazolyl, 1,2,5-thiadiazolyl, phthalimido.

A “4-7 membered saturated heterocyclic group” is a saturated monocyclicring containing 4-7 atoms of which at least one atom is chosen fromnitrogen, sulphur or oxygen, which may, unless otherwise specified, becarbon or nitrogen linked, wherein a —CH₂— group can optionally bereplaced by a —C(O)— and a sulphur atom may be optionally oxidised toform the S-oxides. Examples and suitable values of the term“heterocyclic group” are morpholino, piperidyl, 1,4dioxanyl,1,3-dioxolanyl, 1,2-oxathiolanyl, imidazolidinyl, pyrazolidinyl,piperazinyl, thiazolidinyl, pyrrolidinyl, thiomorpholino,homopiperazinyl and tetrahydropyranyl.

An example of “C₁₋₆alkanoyloxy” is acetoxy. Examples of“C₁₋₆alkoxycarbonyl” include C₁₋₄alkoxycarbonyl, methoxycarbonyl,ethoxycarbonyl, n- and t-butoxycarbonyl. Examples of “C₁₋₆alkoxy”include C₁₋₄alkoxy, C₁₋₃alkoxy, methoxy, ethoxy and propoxy. Examples of“C₁₋₆alkanoylaminol” include formamido, acetamido and propionylamino.Examples of “C₁₋₆alkylS(O)_(a) wherein a is 0 to 2” includeC₁₋₄alkylsulphonyl, methylthio, ethylthio, methylsulphinyl,ethylsulphinyl, mesyl and ethylsulphonyl. Examples of “C₁₋₆alkylS(O)_(r)wherein r is 1 to 2” include methylsulphinyl, ethylsulphinyl, mesyl andethylsulphonyl. Examples of “C₁₋₆alkanoyl” include C₁₋₄alkanoyl,propionyl and acetyl. Examples of “N—C₁₋₆alkylamino” include methylaminoand ethylamino. Examples of “N,N—(C₁₋₆alkyl)₂amino” includedi-N-methylamino, di-(N-ethyl)amino and N-ethyl-N-methylamino. Examplesof “C₂₋₆alkenyl” are vinyl, allyl and 1-propenyl. Examples of“C₂₋₆alkynyl” are ethynyl, 1-propynyl and 2-propynyl. Examples of“N—(C₁₋₆alkyl)sulphamoyl” are N-(methyl)sulphamoyl andN-(ethyl)sulphamoyl. Examples of “N—(C₁₋₆alkyl)₂sulphamoyl” areN,N-(dimethyl)sulphamoyl and N-(methyl)-N-(ethyl)sulphamoyl. Examples of“N—(C₁₋₆alkyl)carbamoyl” are N—(C₁₋₄alkyl)carbamoyl, methylaminocarbonyland ethylaminocarbonyl. Examples of “N,N—(C₁₋₆alkyl)₂carbamoyl” areN,N—(C₁₋₄alkyl)₂carbamoyl, dimethylaminocarbonyl andmethylethylaminocarbonyl. Examples of “C₃₋₈-cycloalkyl” are cyclopropyl,cyclobutyl, cyclopropyl and cyclohexyl. Examples of “(heterocyclicgroup)C₁₋₆alkyl” include pyridylmethyl, 3-morpholinopropyl and2-pyrimid-2-ylethyl. Examples of “C₃₋₈cycloalkylC₁₋₆alkyl” arecyclopropylethyl, cyclobutylmethyl, 2-cyclopropylpropyl andcyclohexylethyl.

A suitable pharmaceutically acceptable salt of a compound of theinvention is, for example, an acid-addition salt of a compound of theinvention which is sufficiently basic, for example, an acid-additionsalt with, for example, an inorganic or organic acid, for examplehydrochloric, hydrobromic, sulphuric, phosphoric, trifluoroacetic,citric or maleic acid. In addition a suitable pharmaceuticallyacceptable salt of a compound of the invention which is sufficientlyacidic is an alkali metal salt, for example a sodium or potassium salt,an alkaline earth metal salt, for example a calcium or magnesium salt,an ammonium salt or a salt with an organic base which affords aphysiologically-acceptable cation, for example a salt with methylamine,dimethylamine, trimethylamine, piperidine, morpholine ortris-(2-hydroxyethyl)amine.

An in vivo hydrolysable ester of a compound of the formula (I)containing carboxy or hydroxy group is, for example, a pharmaceuticallyacceptable ester which is hydrolysed in the human or animal body toproduce the parent acid or alcohol. Suitable pharmaceutically acceptableesters for carboxy include C₁₋₆alkoxymethyl esters for examplemethoxymethyl, C₁₋₆alkanoyloxymethyl esters for examplepivaloyloxymethyl, phthalidyl esters,C₃₋₈cycloalkoxycarbonyloxyC₁₋₆alkyl esters for example1-cyclohexylcarbonyloxyethyl; 1,3-dioxolen-2-onylmethyl esters forexample 5-methyl-1,3-dioxolen-2-onylmethyl; andC₁₋₆alkoxycarbonyloxyethyl esters for example 1-methoxycarbonyloxyethyland may be formed at any carboxy group in the compounds of thisinvention.

An in vivo hydrolysable ester of a compound of the formula (I)containing a hydroxy group includes inorganic esters such as phosphateesters and α-acyloxyalkyl ethers and related compounds which as a resultof the in vivo hydrolysis of the ester breakdown to give the parenthydroxy group. Examples of α-acyloxyalkyl ethers include acetoxymethoxyand 2,2-dimethylpropionyloxy-methoxy. A selection of in vivohydrolysable ester forming groups for hydroxy include alkanoyl, benzoyl,phenylacetyl and substituted benzoyl and phenylacetyl, alkoxycarbonyl(to give alkyl carbonate esters), dialkylcarbamoyl andN-(dialkylaminoethyl)-N-alkylcarbamoyl (to give carbamates),dialkylaminoacetyl and carboxyacetyl. Examples of substituents onbenzoyl include morpholino and piperazino linked from a ring nitrogenatom via a methylene group to the 3- or 4- position of the benzoyl ring.

Some compounds of the formula (I) may have chiral centres and/orgeometric isomeric centres (E- and Z- isomers), and it is to beunderstood that the invention encompasses all such optical,diastereoisomers and geometric isomers that possess CDK inhibitoryactivity.

The invention relates to any and all tautomeric forms of the compoundsof the formula (I) that possess CDK inhibitory activity.

It is also to be understood that certain compounds of the formula (I)can exist in solvated as well as unsolvated forms such as, for example,hydrated forms. It is to be understood that the invention encompassesall such solvated forms which possess CDK inhibitory activity.

Particular values of variable groups are as follows. Such values may beused where appropriate with any of the definitions, claims orembodiments defined hereinbefore or hereinafter.

Ring A is carbocyclyl fused to the imidazole ring.

Ring A is heterocyclyl fused to the imidazole ring.

Ring A is cyclopentyl, cyclohexyl or morpholino fused to the imidazolering.

R¹ is halo.

R¹ is fluoro or chloro.

p is 0-2; wherein the values of R¹ may be the same or different.

p is 0 or 1.

p is 2; wherein the values of R¹ may be the same or different.

p is 1.

p is 0.

R² is a group R^(a)—R^(b)— wherein R^(a) is selected from C₁₋₆alkyloptionally substituted on carbon by one or more R^(g); R^(b) is—N(R^(m))SO₂—; wherein R^(m) is hydrogen; and R^(g) is C₁₋₆alkoxy.

R² is a group R^(a)—R^(b)— wherein R^(a) is selected from ethyloptionally substituted on carbon by one or more R^(g); R^(b) is—N(R^(m))SO₂—; wherein R^(m) is hydrogen; and R^(g) is methoxy orethoxy.

R² is N-(2-methoxyethyl)sulphamoyl or N-(2-ethoxyethyl)sulphamoyl.

q is 0 or 1.

q is 1.

q is 0.

R³ is halo.

R³ is fluoro, chloro or bromo.

R³ is chloro or bromo.

n is 0 or 1.

n is 0.

R⁴ is hydrogen or C₁₋₆alkyl.

R⁴ is hydrogen or methyl.

R⁵ is hydrogen.

m is 1-4; wherein the values of R⁴ may be the same or different.

m is 0 or 1.

m is 0.

m is 1.

Therefore in another aspect of the invention, there is provided acompound of formula (I) (as depicted above) wherein:

Ring A is cyclopentyl, cyclohexyl or morpholino fused to the imidazolering;

p is 0;

R² is a group R^(a)—R^(b)— wherein R^(a) is selected from C₁₋₆alkyloptionally substituted on carbon by one or more R^(g); R^(b) is—N(R^(m))SO₂—; wherein R^(m) is hydrogen; and R^(g) is C₁₋₆alkoxy.

q is 0 or 1;

n is 0;

R⁴ is hydrogen or C₁₋₆alkyl;

R⁵ is hydrogen;

m is 0 or 1;

or a pharmaceutically acceptable salt or an in vivo hydrolysable esterthereof.

Therefore in another aspect of the invention, there is provided acompound of formula (I) (as depicted above) wherein:

Ring A is cyclopentyl, cyclohexyl or morpholino fused to the imidazolering;

p is 0;

R² is N-(2-methoxyethyl)sulphamoyl or N-(2-ethoxyethyl)sulphamoyl;

q is 0 or 1;

n is 0;

R⁴ is hydrogen or methyl;

R⁵ is hydrogen;

m is 0 or 1;

or a pharmaceutically acceptable salt or an in vivo hydrolysable esterthereof.

In another aspect of the invention, preferred compounds of the inventionare any one of the Examples or a pharmaceutically acceptable salt or anin vivo hydrolysable ester thereof

Preferred aspects of the invention are those which relate to thecompound of formula (I) or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention provides a process for preparinga compound of formula (I) or a pharmaceutically acceptable salt or an invivo hydrolysable ester thereof which process (wherein variable groupsare, unless otherwise specified, as defined in formula (I)) comprisesof:

Process a) reaction of a pyrimidine of formula (II):

wherein L is a displaceable group; with an aniline of formula (III):

orProcess b) reacting a compound of formula (IV):

with a compound of formula (V):

wherein T is O or S; R^(x) may be the same or different and is selectedfrom C₁₋₆alkyl; Process c) for compounds of formula (I) where R² issulphamoyl or a group R^(a)—R^(b)— and R^(b) is —NHSO₂—; reacting apyrimidine of formula (VI):

wherein X is a displaceable group; with ammonia or an amine of formula(VII):R^(a)—NH₂  (VII)Process d) for compounds of formula (I); reacting a pyrimidine offormula (VIII)

with a compound of formula (IX):

where Y is a displaceable group;Process e) cyclizing a compound of formula (X):

and thereafter if necessary:

-   i) converting a compound of the formula (I) into another compound of    the formula (I);-   ii) removing any protecting groups;-   iii) forming a pharmaceutically acceptable salt or in vivo    hydrolysable ester.

L is a displaceable group, suitable values for L are for example, ahalogeno or sulphonyloxy group, for example a chloro, bromo,methanesulphonyloxy or toluene-4-sulphonyloxy group.

X is a displaceable group, suitable values for X are for example, afluoro or chloro group. Preferably X is fluoro.

Y is a displaceable group, suitable values for Y are for example, ahalogeno or sulphonyloxy group, for example a bromo, iodo ortrifluoromethanesulphonyloxy group. Preferably Y is iodo.

Specific reaction conditions for the above reactions are as follows.

Process a) Pyrimidines of formula (II) and anilines of formula (III) maybe reacted together:

i) in the presence of a suitable solvent for example a ketone such asacetone or an alcohol such as EtOH or butanol or an aromatic hydrocarbonsuch as toluene or N-methyl pyrrolidine, optionally in the presence of asuitable acid for example an inorganic acid such as hydrochloric acid orsulphuric acid, or an organic acid such as acetic acid or formic acid(or a suitable Lewis acid) and at a temperature in the range of 0° C. toreflux, preferably reflux; orii) under standard Buchwald conditions (for example see J. Am. Chem.Soc., 118, 7215; J. Am. Chem. Soc., 119, 8451; J. Org. Chem., 62, 1568and 6066) for example in the presence of palladium acetate, in asuitable solvent for example an aromatic solvent such as toluene,benzene or xylene, with a suitable base for example an inorganic basesuch as caesium carbonate or an organic base such aspotassium-t-butoxide, in the presence of a suitable ligand such as2,2′-bis(diphenylphosphino)-1,1′-binaphthyl and at a temperature in therange of 25 to 80° C.

Pyrimidines of the formula (II) where L is chloro may be preparedaccording to Scheme 1:

Anilines of formula (III) are commercially available compounds, or theyare known in the literature, or they are prepared by standard processesknown in the art.

Process b) Compounds of formula (IV) and compounds of formula (V) arereacted together in a suitable solvent such as N-methylpyrrolidinone orbutanol at a temperature in the range of 100-200° C., preferably in therange of 150-170° C. The reaction is preferably conducted in thepresence of a suitable base such as, for example, sodium hydride, sodiummethoxide or potassium carbonate.

Compounds of formula (V) may be prepared according to Scheme 2:

Compounds of formula (IV) and (Va) are commercially available compounds,or they are known in the literature, or they are prepared by standardprocesses known in the art.

Process c) Compounds of formula (VI) and amines of formula (VII) may bereacted together in the presence of an inert solvent such asN-methylpyrrolidinone or pyridine, in the presence of a base for examplean inorganic base such as caesium carbonate or in the presence of anorganic base such as excess (VII) and at a temperature in the range of25 to 80° C.

Compounds of formula (VI) (wherein X is chloro) may be preparedaccording to Scheme 3:

Compounds of formula (VIa) may be prepared according to Process a,Process b or Process d wherein q is 0.

Process d) Compounds of formula (VIII) and amines of formula (IX) may bereacted together under standard Buchwald conditions as described inProcess a.

The synthesis of compounds of formula (VIII) is described in Scheme 1.

Compounds of formula (IX) are commercially available compounds, or theyare known in the literature, or they are prepared by standard processesknown in the art

Amines of formula (VI) are commercially available compounds, or they areknown in the literature, or they are prepared by standard processesknown in the art.

Process e) Compounds of formula (X) may be cyclised by treatment withammonium salts, such as ammonium trifluoroacetate or ammonium acetate,at a temperature in the range of 120 to 160° C. either neat or in aninert solvent such as NMP or DMA.

Compounds of formula (X) may be prepared according to Scheme 4:

wherein b is 1-3 and Y is hydrogen or R³.

Compound (Xa) is commercially available.

It will be appreciated that certain of the various ring substituents inthe compounds of the present invention may be introduced by standardaromatic substitution reactions or generated by conventional functionalgroup modifications either prior to or immediately following theprocesses mentioned above, and as such are included in the processaspect of the invention. Such reactions and modifications include, forexample, introduction of a substituent by means of an aromaticsubstitution reaction, reduction of substituents, alkylation ofsubstituents and oxidation of substituents. The reagents and reactionconditions for such procedures are well known in the chemical artParticular examples of aromatic substitution reactions include theintroduction of a nitro group using concentrated nitric acid, theintroduction of an acyl group using, for example, an acyl halide andLewis acid (such as aluminium trichloride) under Friedel Craftsconditions; the introduction of an alkyl group using an alkyl halide andLewis acid (such as aluminium trichloride) under Friedel Craftsconditions; and the introduction of a halogeno group. Particularexamples of modifications include the reduction of a nitro group to anamino group by for example, catalytic hydrogenation with a nickelcatalyst or treatment with iron in the presence of hydrochloric acidwith heating; oxidation of alkylthio to alkylsulphinyl oralkylsulphonyl.

It will also be appreciated that in some of the reactions mentionedherein it may be necessary/desirable to protect any sensitive groups inthe compounds. The instances where protection is necessary or desirableand suitable methods for protection are known to those skilled in theart. Conventional protecting groups may be used in accordance withstandard practice (for illustration see T. W. Green, Protective Groupsin Organic Synthesis, John Wiley and Sons, 1991). Thus, if reactantsinclude groups such as amino, carboxy or hydroxy it may be desirable toprotect the group in some of the reactions mentioned herein.

A suitable protecting group for an amino or alkylamino group is, forexample, an acyl group, for example an alkanoyl group such as acetyl, analkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl ort-butoxycarbonyl group, an arylmethoxycarbonyl group, for examplebenzyloxycarbonyl, or an aroyl group, for example benzoyl. Thedeprotection conditions for the above protecting groups necessarily varywith the choice of protecting group. Thus, for example, an acyl groupsuch as an alkanoyl or alkoxycarbonyl group or an aroyl group may beremoved for example, by hydrolysis with a suitable base such as analkali metal hydroxide, for example lithium or sodium hydroxide.Alternatively an acyl group such as a t-butoxycarbonyl group may beremoved, for example, by treatment with a suitable acid as hydrochloric,sulphuric or phosphoric acid or trifluoroacetic acid and anarylmethoxycarbonyl group such as a benzyloxycarbonyl group may beremoved, for example, by hydrogenation over a catalyst such aspalladium-on-carbon, or by treatment with a Lewis acid for example borontris(trifiuoroacetate). A suitable alternative protecting group for aprimary amino group is, for example, a phthaloyl group which may beremoved by treatment with an alkylamine, for exampledimethylaminopropylamine, or with hydrazine.

A suitable protecting group for a hydroxy group is, for example, an acylgroup, for example an alkanoyl group such as acetyl, an aroyl group, forexample benzoyl, or an arylmethyl group, for example benzyl. Thedeprotection conditions for the above protecting groups will necessarilyvary with the choice of protecting group. Thus, for example, an acylgroup such as an alkanoyl or an aroyl group may be removed, for example,by hydrolysis with a suitable base such as an alkali metal hydroxide,for example lithium or sodium hydroxide. Alternatively an arylmethylgroup such as a benzyl group may be removed, for example, byhydrogenation over a catalyst such as palladium-on-carbon.

A suitable protecting group for a carboxy group is, for example, anesterifying group, for example a methyl or an ethyl group which may beremoved, for example, by hydrolysis with a base such as sodiumhydroxide, or for example a t-butyl group which may be removed, forexample, by treatment with an acid, for example an organic acid such astrifluoroacetic acid, or for example a benzyl group which may beremoved, for example, by hydrogenation over a catalyst such aspalladium-on-carbon.

The protecting groups may be removed at any convenient stage in thesynthesis using conventional techniques well known in the chemical art.

As stated hereinbefore the compounds defined in the present inventionpossesses anti-cell-proliferation activity such as anti-cancer activitywhich is believed to arise from the CDK inhibitory activity of thecompound. These properties may be assessed, for example, using theprocedure set out below:

Assay

The following abbreviations have been used:

-   HEPES is N-[2-Hydroxyethyl]piperazine-N-[2-ethanesulfonic acid]-   DTT is Dithiothreitol-   PMSF is Phenylmethylsulphonyl fluoride

The compounds were tested in an in vitro kinase assay in 96 well formatusing Scintillation Proximity Assay (SPA - obtained from Amersham) formeasuring incorporation of [γ-33-P]-Adenosine Triphosphate into a testsubstrate (GST-Retinoblastoma protein; GST-Rb). In each well was placedthe compound to be tested (diluted in DMSO and water to correctconcentrations) and in control wells either roscovitine as an inhibitorcontrol or DMSO as a positive control.

Approximately 0.2 μl of CDK2/Cyclin E partially-purified enzyme (amountdependent on enzyme activity) diluted in 25 μl incubation buffer wasadded to each well then 20 μl of GST-Rb/ATP/ATP33 mixture (containing0.5 μg GST-Rb and 0.2 μM ATP and 0.14 μCi [γ-33-P]-AdenosineTriphosphate in incubation buffer), and the resulting mixture shakengently, then incubated at room temperature for 60 minutes.

To each well was then added 150 μL stop solution containing (0.8 mg/wellof Protein A-PVT SPA bead (Amersham)), 20 pM/well of Anti-GlutathioneTransferase, Rabbit IgG (obtained from Molecular Probes), 61 mM EDTA and50 mM HEPES pH 7.5 containing 0.05% sodium azide.

The plates were sealed with Topseal-S plate sealers, left for two hoursthen spun at 2500 rpm, 1124 xg., for 5 minutes. The plates were read ona Topcount for 30 seconds per well.

The incubation buffer used to dilute the enzyme and substrate mixescontained 50 mM HEPES pH7.5, 10 mM MnCl₂, 1 mM DTT, 100 μM Sodiumvanadate, 100 μM NaF, 10 mM Sodium Glycerophosphate, BSA (1 mg/mlfinal).

Test Substrate

In this assay only part of the retinoblastoma protein (Science 1987 Mar.13;235(4794):1394-1399; Lee W. H., Bookstein R., Hong F., Young L. J.,Shew J. Y., Lee E. Y.) was used, fused to a GST tag. PCR ofretinoblastoma gene encoding amino acids 379-928 (obtained fromretinoblastoma plasmid ATCC pLRbRNL) was performed, and the sequencecloned into pGEx 2T fusion vector (Smith D. B. and Johnson, K. S. Gene67, 31 (1988); which contained a tac promoter for inducible expression,internal lac I^(q) gene for use in any E. Coli host, and a coding regionfor thrombin cleavage—obtained from Pharmacia Biotech) which was used toamplify amino acids 792-928. This sequence was again cloned into pGEx2T.

The retinoblastoma 792-928 sequence so obtained was expressed in E. Coli(BL21 (DE3) pLysS cells) using standard inducible expression techniques,and purified as follows.

E. coli paste was resuspended in 10 ml/g of NETN buffer (50 mM Tris pH7.5, 120 mM NaCl, 1 mM EDTA, 0.5% v/v NP-40, 1 mM PMSF, 1ug/mlleupeptin, 1ug/ml aprotinin and 1 ug/ml pepstatin) and sonicated for2×45 seconds per 100 ml homogenate. After centrifugation, thesupernatant was loaded onto a 10 ml glutathione Sepharose column(Pharmacia Biotech, Herts, UK), and washed with NETN buffer. Afterwashing with kinase buffer (50 mM HEPES pH 7.5, 10 mM MgC12, 1 mM DTT, 1mM PMSF, 1 ug/ml leupeptin, 1 ug/ml aprotinin and 1 ug/ml pepstatin) theprotein was eluted with 50 mM reduced glutathione in kinase buffer.Fractions containing GST-Rb(792-927) were pooled and dialysed overnightagainst kinase buffer. The final product was analysed by Sodium DodecaSulfate (SDS) PAGE (Polyacrylamide gel) using 8-16% Tris-Glycine gels(Novex, San Diego, USA).

CDK2 and Cyclin E

The open reading frames of CDK2 and Cyclin E were isolated by reversetranscriptase-PCR using HeLa cell and activated T cell mRNA as atemplate and cloned into the insect expression vector pVL1393 (obtainedfrom Invitrogen 1995 catalogue number: V1392-20). CDK2 and cyclin E werethen dually expressed [using a standard virus Baculogold co-infectiontechnique] in the insect SF21 cell system (Spodoptera Frugiperda cellsderived from ovarian tissue of the Fall Army Worm—commerciallyavailable).

Example Production of Cyclin E/CDK2

The following Example provides details of the production of CyclinE/CDK2 in SF21 cells (in TC100+10% FBS(TCS)+0.2% Pluronic) having dualinfection MOI 3 for each virus of Cyclin E & CDK2.

SF21 cells grown in aroller bottle culture to 2.33×10⁶ cells/ml wereused to inoculate 10×500 ml roller bottles at 0.2×10E6 cells/ml. Theroller bottles were incubated on a roller rig at 28° C.

After 3 days (72 hrs.) the cells were counted, and the average from 2bottles found to be 1.86×10E6 cells/ml. (99% viable). The cultures werethen infected with the dual viruses at an MOI 3 for each virus.

The viruses were mixed together before addition to the cultures, and thecultures returned to the roller rig 28° C.

After 2 days (48 hrs.) post infection the 5 Litres of culture washarvested. The total cell count at harvest was 1.58×10E6 cells/ml.(99%viable). The cells were spun out at 2500 rpm, 30 mins., 4° C. in HeraeusOmnifuge 2.0 RS in 250 ml. lots. The supernatant was discarded.

Partial Co-Purification of Cdk2 and Cyclin E

Sf21 cells were resuspended in lysis buffer (50 mM Tris pH 8.2, 10 mMMgCl₂, 1 mM DTT, 10 mM glycerophosphate, 0.1 mM sodium orthovanadate,0.1 mM NaF, 1 mM PMSF, 1 ug/ml leupeptin and 1 ug/ml aprotinin) andhomogenised for 2 minutes in a 10 ml Dounce homgeniser. Aftercentrifugation, the supernatant was loaded onto a Poros HQ/M 1.4/100anion exchange column (PE Biosystems, Hertford, UK). Cdk2 and Cyclin Ewere coeluted at the beginning of a 0-1M NaCl gradient (run in lysisbuffer minus protease inhibitors) over 20 column volumes. Co-elution waschecked by western blot using both anti-Cdk2 and anti-Cyclin Eantibodies (Santa Cruz Biotechnology, Calif., US).

By analogy, assays designed to assess inhibition of CDK1 and CDK4 may beconstructed. CDK2 (EMBL Accession No. X62071) may be used together withCyclin A or Cyclin E (see EMBL Accession No. M73812), and furtherdetails for such assays are contained in PCT International PublicationNo. WO99/21845, the relevant Biochemical & Biological Evaluationsections of which are hereby incorporated by reference.

Although the pharmacological properties of the compounds of the formula(I) vary with structural change, in general activity possessed bycompounds of the formula (I) may be demonstrated at IC₅₀ concentrationsor doses in the range 250 μM to 1 nM.

The following IC₅₀s were measured in the above assay.

Example No IC₅₀ 2 21 nM 5 42 nM 9 13 nM

The in vivo activity of the compounds of the present invention may beassessed by standard techniques, for example by measuring inhibition ofcell growth and assessing cytotoxicity.

Inhibition of cell growth may be measured by staining cells withSulforhodamine B (SRB), a fluorescent dye that stains proteins andtherefore gives an estimation of amount of protein (i.e. cells) in awell (see Boyd, M. R.(1989) Status of the NCI preclinical antitumourdrug discovery screen. Prin. Prac Oncol 10:1-12). Thus, the followingdetails are provided of measuring inhibition of cell growth:—

Cells were plated in appropriate medium in a volume of 100 μl in 96 wellplates; media was Dulbecco's Modified Eagle media for MCF-7, SK-UT-1Band SK-UT-1. The cells were allowed to attach overnight, then inhibitorcompounds were added at various concentrations in a maximumconcentration of 1% DMSO (v/v). A control plate was assayed to give avalue for cells before dosing. Cells were incubated at 37° C., (5% CO₂)for three days.

At the end of three days TCA was added to the plates to a finalconcentration of 16% (v/v). Plates were then incubated at 4° C. for 1hour, the supernatant removed and the plates washed in tap water. Afterdrying, 100 μl SRB dye (0.4% SRB in 1% acetic acid) was added for 30minutes at 37° C. Excess SRB was removed and the plates washed in 1%acetic acid. The SRB bound to protein was solubilised in 10 mM TrispH7.5 and shaken for 30 minutes at room temperature. The ODs were readat 540 nm, and the concentration of inhibitor causing 50% inhibition ofgrowth was determined from a semi-log plot of inhibitor concentrationversus absorbance. The concentration of compound that reduced theoptical density to below that obtained when the cells were plated at thestart of the experiment gave the value for toxicity.

Typical IC₅₀ values for compounds of the invention when tested in theSRB assay are in the range 1 mM to 1 nM.

According to a further aspect of the invention there is provided apharmaceutical composition which comprises a pyrimidine derivative ofthe formula (I), or a pharmaceutically acceptable salt or in vivohydrolysable ester thereof, as defined hereinbefore in association witha pharmaceutically-acceptable diluent or carrier.

The composition may be in a form suitable for oral administration, forexample as a tablet or capsule, for parenteral injection (includingintravenous, subcutaneous, intramuscular, intravascular or infusion) asa sterile solution, suspension or emulsion, for topical administrationas an ointment or cream or for rectal administration as a suppository.

In general the above compositions may be prepared in a conventionalmanner using conventional excipients.

The compound of formula (I) will normally be administered to awarm-blooded animal at a unit dose within the range 5-5000 mg per squaremeter body area of the animal, i.e. approximately 0.1-100 mg/kg, andthis normally provides a therapeutically-effective dose. A unit doseform such as a tablet or capsule will usually contain, for example 1-250mg of active ingredient. Preferably a daily dose in the range of 1-50mg/kg is employed. However the daily dose will necessarily be varieddepending upon the host treated, the particular route of administration,and the severity of the illness being treated. Accordingly the optimumdosage may be determined by the practitioner who is treating anyparticular patient.

According to a further aspect of the present invention there is provideda compound of the formula (I), or a pharmaceutically acceptable salt orin vivo hydrolysable ester thereof, as defined hereinbefore for use in amethod of treatment of the human or animal body by therapy.

We have found that the compounds defined in the present invention, or apharmaceutically acceptable salt or in vivo hydrolysable ester thereof,are effective cell cycle inhibitors (anti-cell proliferation agents),which property is believed to arise from their CDK inhibitoryproperties. Accordingly the compounds of the present invention areexpected to be useful in the treatment of diseases or medical conditionsmediated alone or in part by CDK enzymes, i.e. the compounds may be usedto produce a CDK inhibitory effect in a warm-blooded animal in need ofsuch treatment. Thus the compounds of the present invention provide amethod for treating the proliferation of malignant cells characterisedby inhibition of CDK enzymes, i.e. the compounds may be used to producean anti-proliferative effect mediated alone or in part by the inhibitionof CDKs. Such a compound of the invention is expected to possess a widerange of anti-cancer properties as CDKs have been implicated in manycommon human cancers such as leukaemia and breast, lung, colon, rectal,stomach, prostate, bladder, pancreas and ovarian cancer. Thus it isexpected that a compound of the invention will possess anti-canceractivity against these cancers. It is in addition expected that acompound of the present invention will possess activity against a rangeof leukaemias, lymphoid malignancies and solid tumours such ascarcinomas and sarcomas in tissues such as the liver, kidney, prostateand pancreas. In particular such compounds of the invention are expectedto slow advantageously the growth of primary and recurrent solid tumoursof, for example, the colon, breast, prostate, lungs and skin. Moreparticularly such compounds of the invention, or a pharmaceuticallyacceptable salt or in vivo hydrolysable ester thereof, are expected toinhibit the growth of those primary and recurrent solid tumours whichare associated with CDKs, especially those tumours which aresignificantly dependent on CDKs for their growth and spread, includingfor example, certain tumours of the colon, breast, prostate, lung, vulvaand skin.

It is further expected that a compound of the present invention willpossess activity against other cell-proliferation diseases in a widerange of other disease states including leukaemias, fibroproliferativeand differentiative disorders, psoriasis, rheumatoid arthritis, Kaposi'ssarcoma, haemangioma, acute and chronic nephropathies, atheroma,atherosclerosis, arterial restenosis, autoimmune diseases, acute andchronic inflammation, bone diseases and ocular diseases with retinalvessel proliferation.

Thus according to this aspect of the invention there is provided acompound of the formula (I), or a pharmaceutically acceptable salt or invivo hydrolysable ester thereof, as defined hereinbefore for use as amedicament; and the use of a compound of the formula (I), or apharmaceutically acceptable salt or in vivo hydrolysable ester thereof,as defined hereinbefore in the manufacture of a medicament for use inthe production of a cell cycle inhibitory (anti-cell-proliferation)effect in a warm-blooded animal such as man. Particularly, an inhibitoryeffect is produced by preventing entry into or progression through the Sphase by inhibition of CDK2 and CDK4, especially CDK2, and M phase byinhibition of CDK1.

According to a further feature of the invention, there is provided acompound of the formula (I), or a pharmaceutically acceptable salt or invivo hydrolysable ester thereof, as defined herein before in themanufacture of a medicament for use in the treatment of cancers (solidtumours and leukaemias), fibroproliferative and differentiativedisorders, psoriasis, rheumatoid arthritis, Kaposi's sarcoma,haemangioma, acute and chronic nephropathies, atheroma, atherosclerosis,arterial restenosis, autoimmune diseases, acute and chronicinflammation, bone diseases and ocular diseases with retinal vesselproliferation, particularly in the treatment of cancers.

According to a further feature of this aspect of the invention there isprovided a method for producing a cell cycle inhibitory(anti-cell-proliferation) effect in a warm-blooded animal, such as man,in need of such treatment which comprises administering to said animalan effective amount of a compound as defined immediately above.Particularly, an inhibitory effect is produced by preventing entry intoor progression through the S phase by inhibition of CDK2 and CDK4,especially CDK2, and M phase by inhibition of CDK1.

According to a further feature of this aspect of the invention there isprovided a method for producing a cell cycle inhibitory(anti-cell-proliferation) effect in a warm-blooded animal, such as man,in need of such treatment which comprises administering to said animalan effective amount of a compound of formula (I) or a pharmaceuticallyacceptable salt or in vivo hydrolysable ester thereof as defined hereinbefore. Particularly, an inhibitory effect is produced by preventingentry into or progression through the S phase by inhibition of CDK2 andCDK4, especially CDK2, and M phase by inhibition of CDK1.

According to an additional feature of this aspect of the invention thereis provided a method of treating cancers (solid tumours and leukaemias),fibroproliferative and differentiative disorders, psoriasis, rheumatoidarritis, Kaposi's sarcoma, haemangioma, acute and chronic nephropathies,atheroma, atherosclerosis, arterial restenosis, autoimmune diseases,acute and chronic inflammafion, bone diseases and ocular diseases withretinal vessel proliferation, in a warm-blooded animal, such as man, inneed of such treatment which comprises administering to said animal aneffective amount of a compound of formula (I) or a pharmaceuticallyacceptable salt or int vivo hydrolysable ester thereof as defined hereinbefore.

Particularly there is provided a method of treating cancer in awarm-blooded animal, such as man, in need of such treatment whichcomprises administering to said animal an effective amount of a compoundof formula (I) or a pharmaceutically acceptable salt or in vivohydrolysable ester thereof as defined herein before.

In a further aspect of the invention there is provided a pharmaceuticalcomposition which comprises a compound of the formula (I), or apharmaceutically acceptable salt or in vivo hydrolysable ester thereof,as defined herein before in association with apharmaceutically-acceptable diluent or carrier for use in the productionof a cell cycle inhibitory (anti-cell-proliferation) effect in awarm-blooded animal such as man.

In a further aspect of the invention there is provided a pharmaceuticalcomposition which comprises a compound of the formula (I), or apharmaceutically acceptable salt or in vivo hydrolysable ester thereof,as defined herein before in association with apharmaceutically-acceptable diluent or carrier for use in the treatmentof cancers (solid tumours and leukaemias), fibroproliferative anddifferentiative disorders, psoriasis, rheumatoid arthritis, Kaposi'ssarcoma, haemangioma, acute and chronic nephropathies, atheroma,atherosclerosis, arterial restenosis, autoimmune diseases, acute andchronic inflammation, bone diseases and ocular diseases with retinalvessel proliferation, in a warm-blooded animal such as man.

In a further aspect of the invention there is provided a pharmaceuticalcomposition which comprises a compound of the formula (I), or apharmaceutically acceptable salt or in vivo hydrolysable ester thereof,as defined herein before in association with apharmaceutically-acceptable diluent or carrier for use in the treatmentof cancer in a warm-blooded animal such as man.

Preventing cells from entering DNA synthesis by inhibition of essentialS-phase initiating activities such as CDK2 initiation may also be usefulin protecting normal cells of the body from toxicity of cycle-specificpharmaceutical agents. Inhibition of CDK2 or 4 will prevent progressioninto the cell cycle in normal cells which could limit the toxicity ofcycle-specific pharmaceutical agents which act in S-phase, G2 ormitosis. Such protection may result in the prevention of hair lossnormally associated with these agents.

Therefore in a further aspect of the invention there is provided acompound of formula (I) as defined above or a pharmaceuticallyacceptable salt or in vivo hydrolysable ester thereof for use as a cellprotective agent.

Therefore in a further aspect of the invention there is provided acompound of formula (I) as defined above or a pharmaceuticallyacceptable salt or in vivo hydrolysable ester thereof for use inpreventing hair loss arising from the treatment of malignant conditionswith pharmaceutical agents.

Examples of pharmaceutical agents for treating malignant conditions thatare known to cause hair loss include alkylating agents such asifosfamide and cyclophosphamide; antimetabolites such as methotrexate,5-fluorouracil, gemcitabine and cytarabine; vinca alkaloids andanalogues such as vincristine, vinbalstine, vindesine, vinorelbine;taxanes such as paclitaxel and docetaxel; topoisomerase I inhibitorssuch as irintotecan and topotecan; cytotoxic antibiotics such asdoxorubicin, daunorubicin, mitoxantrone, actinomycin-D and mitomycin;and others such as etoposide and tretinoin.

In another aspect of the invention, the compound of formula (I), or apharmaceutically acceptable salt or in vivo hydrolysable ester thereof,may be administered in association with a one or more of the abovepharmaceutical agents. In this instance the compound of formula (I) maybe administered by systemic or non systemic means. Particularly thecompound of formula (I) my may administered by non-systemic means, forexample topical administration.

Therefore in an additional feature of the invention, there is provided amethod of preventing hair loss during treatment for one or moremalignant conditions with pharmaceutical agents, in a warm-bloodedanimal, such as man, which comprises administering to said animal aneffective amount of a compound of formula (I), or a pharmaceuticallyacceptable salt or in vivo hydrolysable ester thereof.

In an additional feature of the invention, there is provided a method ofpreventing hair loss during treatment for one or more malignantconditions with pharmaceutical agents, in a warm-blooded animal, such asman, which comprises administering to said animal an effective amount ofa compound of formula (I), or a pharmaceutically acceptable salt or invivo hydrolysable ester thereof in simultaneous, sequential or separateadministration with an effective amount of said pharmaceutical agent.

According to a further aspect of the invention there is provided apharmaceutical composition for use in preventing hair loss arising fromthe treatment of malignant conditions with pharmaceutical agents whichcomprises a compound of formula (I), or a pharmaceutically acceptablesalt or in vivo hydrolysable ester thereof, and said pharmaceuticalagent, in association with a pharmaceutically acceptable diluent orcarrier.

According to a further aspect of the present invention there is provideda kit comprising a compound of formula (I), or a pharmaceuticallyacceptable salt or in vivo hydrolysable ester thereof and apharmaceutical agent for treating malignant conditions that is known tocause hair loss.

According to a further aspect of the present invention there is provideda kit comprising:

-   a) a compound of formula (I), or a pharmaceutically acceptable salt    or in vivo hydrolysable ester thereof, in a first unit dosage form;-   b) a pharmaceutical agent for treating malignant conditions that is    known to cause hair loss; in a second unit dosage form; and-   c) container means for containing said first and second dosage    forms.

According to another feature of the invention there is provided the useof a compound of the formula (I), or a pharmaceutically acceptable saltor in vivo hydrolysable ester thereof, in the manufacture of amedicament for the prevention of hair loss during treatment of malignantconditions with pharmaceutical agents.

According to a further aspect of the present invention there is provideda combination treatment for the prevention of hair loss comprising theadministration of an effective amount of a compound of the formula (I),or a pharmaceutically acceptable salt or in vivo hydrolysable esterthereof, optionally together with a pharmaceutically acceptable diluentor carrier, with the simultaneous, sequential or separate administrationof an effective amount of a pharmaceutical agent for treatment ofmalignant conditions to a warm-blooded animal, such as man.

As stated above the size of the dose required for the therapeutic orprophylactic treatment of a particular cell-proliferation disease willnecessarily be varied depending on the host treated, the route ofadministration and the severity of the illness being treated. A unitdose in the range, for example, 1-100 mg/kg, preferably 1-50 mg/kg isenvisaged.

The CDK inhibitory activity defined hereinbefore may be applied as asole therapy or may involve, in addition to a compound of the invention,one or more other substances and/or treatments. Such conjoint treatmentmay be achieved by way of the simultaneous, sequential or separateadministration of the individual components of the treatment. In thefield of medical oncology it is normal practice to use a combination ofdifferent forms of treatment to treat each patient with cancer. Inmedical oncology the other component(s) of such conjoint treatment inaddition to the cell cycle inhibitory treatment defined hereinbefore maybe: surgery, radiotherapy or chemotherapy. Such chemotherapy may coverthree main categories of therapeutic agent:

-   (i) other cell cycle inhibitory agents that work by the same or    different mechanisms from those defined hereinbefore;-   (ii) cytostatic agents such as antioestrogens (for example    tamoxifen,toremifene, raloxifene, droloxifene, iodoxyfene),    progestogens (for example megestrol acetate), aromatase inhibitors    (for example anastrozole, letrazole, vorazole, exemestane),    antiprogestogens, antiandrogens (for example flutamide, nilutamide,    bicalutamide, cyproterone acetate), LHRH agonists and antagonists    (for example goserelin acetate, luprolide), inhibitors of    testosterone 5α-dihydroreductase (for example finasteride),    anti-invasion agents (for example metalloproteinase inhibitors like    marimastat and inhibitors of urokinase plasminogen activator    receptor function) and inhibitors of growth factor function, (such    growth factors include for example platelet derived growth factor    and hepatocyte growth factor such inhibitors include growth factor    antibodies, growth factor receptor antibodies, tyrosine kinase    inhibitors and serine/threonine kinase inhibitors); and-   (iii) antiproliferative/antineoplastic drugs and combinations    thereof, as used in medical oncology, such as antimetabolites (for    example antifolates like methotrexate, fluoropyrimidines like    5-fluorouracil, purine and adenosine analogues, cytosine    arabinoside); antitumour antibiotics (for example anthracyclines    like doxorubicin, daunomycin, epirubicin and idarubicin,    mitomycin-C, dactinomycin, mithramycin); platinum derivatives (for    example cisplatin, carboplatin); alkylating agents (for example    nitrogen mustard, melphalan, chlorambucil, busulphan,    cyclophosphamide, ifosfamide, nitrosoureas, thiotepa); antimitotic    agents (for example vinca alkaloids like vincristine and taxoids    like taxol, taxotere); topoisomerase inhibitors (for example    epipodophyllotoxins like etoposide and teniposide, amsacrine,    topotecan). According to this aspect of the invention there is    provided a pharmaceutical product comprising a compound of the    formula (I) as defined hereinbefore and an additional anti-tumour    substance as defined hereinbefore for the conjoint treatment of    cancer.

In addition to their use in therapeutic medicine, the compounds offormula (I) and their pharmaceutically acceptable salts are also usefulas pharmacological tools in the development and standardisation of invitro and in vivo test systems for the evaluation of the effects ofinhibitors of cell cycle activity in laboratory animals such as cats,dogs, rabbits, monkeys, rats and mice, as part of the search for newtherapeutic agents.

In the above other pharmaceutical composition, process, method, use andmedicament manufacture features, the alternative and preferredembodiments of the compounds of the invention described herein alsoapply.

EXAMPLES

The invention will now be illustrated by the following non limitingexamples in which, unless stated otherwise:

-   (i) temperatures are given in degrees Celsius (° C.); operations    were carried out at room or ambient temperature, that is, at a    temperature in the range of 18-25° C;-   (ii) organic solutions were dried over anhydrous magnesium sulphate;    evaporation of solvent was carried out using a rotary evaporator    under reduced pressure (600-4000 Pascals; 4.5-30 mmHg) with a bath    temperature of up to 60° C.;-   (iii) chromatography means flash chromatography on silica gel; thin    layer chromatography (TLC) was carried out on silica gel plates;-   (iv) in general, the course of reactions was followed by TLC and    reaction times are given for illustration only;-   (v) final products had satisfactory proton nuclear magnetic    resonance (NMR) spectra and/or mass spectral data;-   (vi) yields are given for illustration only and are not necessarily    those which can be obtained by diligent process development;    preparations were repeated if more material was required;-   (vii) when given, NMR data is in the form of delta values for major    diagnostic protons, given in parts per million (ppm) relative to    tetramethylsilane (TMS) as an internal standard, determined at 300    MHz using perdeuterio dimethyl sulphoxide (DMSO-d₆) as solvent    unless otherwise indicated;-   (viii) chemical symbols have their usual meanings; SI units and    symbols are used;-   (ix) solvent ratios are given in volume:volume (v/v) terms; and-   (x) mass spectra were run with an electron energy of 70 electron    volts in the chemical ionization (CI) mode using a direct exposure    probe; where indicated ionization was effected by electron impact    (El), fast atom bombardment (FAB) or electrospray (ESP); values for    m/z are given; generally, only ions which indicate the parent mass    are reported; and unless otherwise stated, the mass ion quoted is    (MH)⁺;-   (xi) unless stated otherwise compounds containing an asymmetrically    substituted carbon and/or sulphur atom have not been resolved;-   (xii) where a synthesis is described as being analogous to that    described in a previous example the amounts used are the millimolar    ratio equivalents to those used in the previous example; and-   (xvi) the following abbreviations have been used:

THF tetrahydrofuran;

DMF N,N-dimethylformamide;

EtOAc ethyl acetate;

MeOH methanol;

EtOH ethanol;

DCM dichloromethane; and

DMSO dimethylsulphoxide.

Example 12-Anilino-4-(6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)pyrimidine

A mixture of2-anilino-4-[1-(pyrrolid-2-on-1-yl)-2-(dimethylamino)vinyl]pyrimidine(Method 6; 1.85 g, 5.72 mmol) and ammonium trifluoroacetate (7.5 g, 57.2mmol) in dry N-methylpyrrolidinone was stirred and heated at 140° C.under nitrogen for 18 hours and then at 150° C. for 10 hours. Themixture was allowed to cool and the solvent removed by evaporation. Theresidue was partitioned between water and DCM and insolubles removed byfiltration. The biphasic mixture was separated and the organic layerdried (Na₂SO₄) and the solvent removed by evaporation. The residue waspurified by chromatography eluting with DCM/MeOH (98:2 and then96.5:3.5). The purified product was triturated with diethyl ether,filtered and dried to give the title compound (120 mg, 8%) as a yellowsolid. NMR: 2.55 (m, 2H), 2.78 (t, 2H), 4.33 (t, 2H), 6.95 (t, 1H), 7.08(d, 1H), 7.27 (t, 2H), 7.7(d, 2H), 7.75 (s, 1H), 8.35 (d, 1H), 9.38 (s,1H): m/z 278.

Examples 2-5

The following compounds were prepared by the procedure of Example 1using the appropriate starting materials (wherein “*” represents thepoint of attachment to the pyrimidine ring):

Ex X NMR m/z SM  2¹

1.87(m, 4H), 2.83(t, 2H), 4.42(t, 2H), 6.95(t, 1H), 7.08(d, 1H), 7.3(t,2H), 7.68(m, 3H), 8.35(d, 1H), 9.37(s, 1H) 292 Meth 7 3

1.23(d, 3H), 2.23(m, 1H), 2.77(m, 2H), 2.9(m, 1H), 5.08(m, 1H), 6.97(t,1H), 7.1(d, 1H), 7.3(t, 2H), 7.67(d, 2H), 7.77(s, 1H), 8.35(d, 1H),9.35(s, 1H) 292 Meth 8 4

1.17(d, 3H), 1.9(m, 4H), 2.8(m, 2H), 5.53(m, 1H), 6.95(t, 1H), 7.08(d,1H), 7.37(t, 2H), 7.65(m, 2H), 8.33(d, 1H), 9.33(s, 1H) 306 Meth 9 5

4.03(t, 2H), 4.47(t, 2H), 4.8(s, 2H), 6.95(t, 1H), 7.12(d, 1H), 7.27(t,2H), 7.67(d, 2H), 7.77(s, 1H), 8.38(d, 1H), 9.4(s, 1H) 294 Meth 10¹Purified by chromatography eluting with DCM/MeOH(95:5).

Example 62-[4-(N-(2-Methoxyethyl)sulphamoyl)anilino]-4-(6,7-dihydro-5H-pyrrolo[,1,2-a]imidazol-3-yl)pyrimidine

Chlorosulphonic acid (143 μl, 2.16 mmol) was added to a stirredsuspension of2-anilino-4-(6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)pyrimidine(Example 1; 150 mg, 0.54 mmol) in thionyl chloride (3.0 ml) at 0° C. Themixture was stirred at 0° C. for 10 minutes and then heated at refluxfor 80 minutes. The mixture was allowed to cool and the excess thionylchloride removed by evaporation. A solution of 2-methoxyethylamine(0.756 ml, 8.1 mmol) in MeOH (1.5 ml) was added to the residue and themixture stirred at ambient temperature for 24 hours. The volatiles wereremoved by evaporation and the residue suspended in distilled water andstirred for 1 hour. The resulting solid was collected by filtration,washed with water and dried to give the title compound (182 mg, 81%) asa pale brown solid. NMR: 2.58 (m, 2H), 2.83 (m, 4H), 3.18 (s, 3H), 3.3(t, 2H), 4.4 (t, 2H), 7.2 (d, 1H), 7.47 (s, 1H), 7.7 (d, 2H), 7.8 (s.1H), 7.92 (d, 2H), 8.43 (d, 1H), 9.83 (s, 1H): m/z 415.

Examples 7-9

The following compounds were prepared by the procedure of Example 6using the appropriate starting materials.

Ex R X NMR m/z SM 7 Me CH₂ 1.9(m, 4H), 2.86(m, 4H), 3.17(s, 3H), 429 Ex2 3.3(t, 2H), 4.47(t, 2H), 7.2(d, 1H), 7.45(t, 1H), 7.7(m, 3H), 7.9(d,2H), 8.4(d, 1H), 9.83(s, 1H) 8 Et CH₂ 1.0(t, 3H), 1.83(m, 4H), 2.78(m,4H), 443 Ex 2 3.25(m, 4H), 4.4(t, 2H), 7.13(d, 1H), 7.40(t, 1H), 7.65(m,3H), 7.83(d, 2H), 8.37(d, 1H), 9.80(s, 1H) 9 Me O 2.9(t, 2H), 3.17(s,3H), 3.3(t, 2H), 4.04(t, 431 Ex 5 2H), 4.53(t., 2H), 4.83(s, 2H),7.23(d, 1H), 7.43(s, 1H), 7.72(d, 2H), 7.8(s, 1H), 7.88(d, 2H), 8.45(d,1H), 9.86(s, 2H):

Example 102-Anilino-4-(8-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyridin-3-yl)pyrimidine

n-Butyllithium (1.2 ml of a 1.6M solution in hexane, 1.92 mmol) wasadded slowly to a stirred solution of2-anilino-4-(5,6,7,8-tetrahydroimidazo[1,2-a]pyridin-3-yl)pyrimidine(Example 2; 200 mg, 0.69 mmol) in dry THF (10 ml) cooled to −78° C.under nitrogen making sure that the reaction temperature remained below−65° C. The mixture was stirred at −70° C. for 15 minutes, theniodomethane (45 μl, 0.72 mmol) was added and the reaction stirred at−70° C. for 10 minutes and then allowed to warm and stirred at ambienttemperature for 20 hours. The reaction mixture was partitioned betweenEtOAc and water and the layers separated. The organic layer was washedwith saturated brine, dried (Na₂SO₄) and the volatiles removed byevaporation. The residue was purified by chromatography eluting withDCM/MeOH (98:2). The purified product was twice triturated with diethylether, collected by filtration and dried to give the title compound (33mg, 16%) as a pale yellow solid. NMR: 1.37 (d, 3H), 1.57 (m, 1H), 1.92(m, 1H), 2.07 (m, 2H), 2.97 (m, 1H), 4.25 (m, 1H), 4.65 (m, 1H), 7.02(t, 2H), 7.15 (d, 1H), 7.33 (t, 2H), 7.72 (m, 3H), 8.38 (d, 1H), 9.42(s. 1H); m/z 306.

Example 112-Anilino-4(8-methyl-5.6-dihydro-8H-imidazo[2,1-c][1,4]oxazin-3-yl)pyrimidine

By an analogous process to Example 10,2-Anilino-4-(5,6-dihydro-8H-imidazo[2,1-c][1,4]oxazin-3-yl)pyrimidine(Example 5; 163 mg, 0.556 mmol) was treated with n-butyllithium (0.73 mlof a 1.6M solution in hexane, 1.168 mmol) and iodomethane (36 μl, 0.583mmol) to give the title compound (67 mg, 39%) as a pale yellow solid.NMR: 1.5 (d, 3H), 3.9 (m, 1H), 4.18 (m, 1H), 4.32 (m, 1H), 4.63 (m, 1H),4.87 (q, 1H), 6.95 (t, 1H), 7.13 (d, 1H), 7.3 (t, 2H), 7.67 (d, 2H),7.75 (s, 1H), 8.37 (d, 1H), 9.4 (s, 1H); m/z 308.

Preparation of Starting Materials

The starting materials for the above Examples are either commerciallyavailable or are readily prepared by standard methods from knownmaterials. For example the following reactions are illustrations but notlimitations of the preparation of some of the starting materials used inthe above reactions.

Method 1

1,1-Dimethoxy-4-dimethylaminobut-3-en-2-one

1,1-Dimethoxypropan-2-one (57.9 g, 0.459 mol) was added to dimethylformamide dimethyl acetal (54.6 g, 0.459 mol) and the mixture stirredand heated at 100° C. for 18 hours. The reaction mixture was allowed tocool and the volatiles removed by evaporation give the itle compound (79g) as a red oil. NMR: (CDCl₃): 2.90 (d, 3H), 3.10 (d, 3H), 3.40 (s, 6H),4.56 (s, 1H), 5.34 (1H, d), 7.74 (d, 1H).

Method 2

2-Anilino-4-(dimethoxymethyl)pyrimidine

A mixture of 1,1-dimethoxy-4-dimethylaminobut-3-en-2-one (Method 1; 20.0g, 0.116 mol) and phenyl guanidine bicarbonate (25 g, 0.126 mol) inanhydrous dimethylacetamide (350 ml) was stirred and heated at 130° C.under nitrogen for 18 hours. The reaction mixture was allowed to cooland the solvent removed by evaporation. The residue was dissolved inEtOAc and the solution washed with water (×5), saturated sodium chloride(×2), dried and evaporated to give the title compound (24.7 g, 88%) as adark brown oil. NMR: (CDCl₃): 3.42 (s, 6H), 5.20 (s, 1H), 6.94 (d, 1H),7.04 (t, 1H), 7.31 (m, 3H), 7.64 (d, 2H), 8.47 (d, 1H), m/z: 246.

Method 3

2-Anilinopyrimidin-4-carbaldehyde

A mixture of 2-anilino-4-(dimethoxymethyl)pyrimidine (Method 2; 26.5 g,0.108 mol) and 3M hydrochloric acid (108 ml, 0.324 mol) was stirred andheated at 50° C. for 18 hours. The reaction mixture was allowed to cooland the pH of the solution adjusted to 9 by careful addition of solidsodium carbonate. The mixture was extracted with EtOAc (5×100 ml). Thecombined extracts were washed with water (2×100 ml), dried (Na₂SO₄) andevaporated to give the title compound (16.2 g, 78%) as a brown solid.NMR: (CDCl₃): 7.10 (t, 1H), 7.20 (d, 1H), 7.40 (t, 3H), 7.68 (d, 21),8.64 (d, 1H), 9.90 (s, 1H), m/z 232 [MH+MeOH]⁺.

Method 4

2-Anilino-4-(pyrrolid-2-on-1-ylmethyl)pyrimidine

Triethylamine (1.52 ml, 11 mmol) was added to a mixture of2-anilinopyrimidin-4-carbaldehyde (Method 3; 1.99 g, 10 mmol) and ethyl4-aminobutyrate hydrochloride (1.84 g, 11 mmol) in MeOH (100 ml) and themixture stirred at ambient temperature under nitrogen for 18 hours. Theinsolubles were removed by filtration and the volatiles removed from thefiltrate by evaporation. The residue was suspended in EtOAc and furtherinsoluble material removed by filtration and the filtrate evaporated.The residue was suspended in EtOH (200 ml) and stirred under nitrogen.Sodium borohydride (0.456 g, 12 mmol) was added in several portions andthe reaction mixture then stirred at ambient temperature for 96 hours.The solvent was removed by evaporation, the residue dissolved in EtOAc(150 ml) and the solution washed with saturated brine, dried and thevolatiles evaporated to give the title compound (2.2 g, 88%) as a yellowgum. NMR: (CDCl₃) 2.12 (m, 2H), 2,47 (t, 2H), 3.45 (t, 2H), 4.44 (s,2H), 6.62 (d, 1H), 7.04 (t, 1H), 7.32 (m, 3H), 7.61 (d, 2H), 8.35 (d,1H), m/z 269.

Method 5

2-Anilino-4-(piperidin-2-on-1-ylmethyl)pyrimidine

By an analogous process to Method 4, 2-anilinopyrimidin-4-carbaldehyde(Method 3; 3.24, 16.2 mmol) was treated with methyl 5-aminopentanoatehydrochloride (3.0 g, 17.9 mmol) to give the title compound (2.23 g,51%) as a pale yellow solid. NMR: 1.85 (m, 4H), 2.5 (s, 2H), 3.4 (s,2H), 4.58 (s, 2H), 6.63 (d, 1H), 7.05 (t, 1H), 7.3 (m, 3H), 7.62 (d,2H), 8.34 (d, 1H); m/z 305.

Method 6

2-Anilino-4[1-(pyrrolid-2-on-1-yl)-2-(dimethylamino)vinyl]pyrimidine

A mixture of 2-anilino-4-(pyrrolid-2-on-1-ylmethyl)pyrimidine (Method 4;960 mg, 3.6 mmol) and tris-dimethylaminomethane (1.04 g, 7.2 mmol) indry DMF (15 ml) was stirred and heated at 130° C. under nitrogen for 6hours. The reaction mixture was allowed to cool and the solvent removedby evaporation. The residue was triturated with a mixture of ether anddistilled water and the solid product collected by filtration, washedwith water and ether, and dried to give the title compound (745 mg, 81%)as a tan solid. NMR: (CDCl₃): 2.11 (m, 2H), 2,45 (m, 2H), 3.00 (s, 6H),3.38 (m, 1H), 3.72 (m, 1H), 6.20 (d, 1H), 7.00 (t, 1H), 7.08 (s, 1H),7.27 (m, 2H), 7.58 (m, 2H), 7.7 (s, 1H), 8.11(d, 1H); m/z 324.

Methods 7-10

The following compounds were prepared by the procedure of Method 6 usingthe appropriate starting materials (wherein “*” represents the point ofattachment).

Meth Ring A NMR m/z SM  7

1.9(m, 4H), 2.52(m, 2H), 3.0(s, 6H), 3.37(m, 2H), 6.22(d, 1H), 6.96(m,1H), 7.3(m, 4H), 7.58(d, 2H), 7.65(s, 1H), 8.07(d, 1H) 338 Meth 5  8¹

1.1(d, 3H), 1.7(m, 1H), 2.37(m, 3H), 3.83(m, 1H), 6.33(d, 1H), 6.92(t,1H), 7.25(t, 2H), 7.7(m, 3H), 8.05(d, 1H), 8.6(s, 1H) 338 Meth 15  9¹

1.17(d, 3H), 1.83(m, 4H), 2.4(m, 2H), 3.0(s, 6H), 3.7(q, 1H), 6.33(d,1H), 6.93(t, 1H), 7.26(t, 2H), 7.63(s, 1H), 7.73(d, 1H), 8.05(d, 1H),8.4(s, 1H) 352 Meth 18 10²

3.0(s, 6H), 3.38(m, 1H), 3.47(m, 1H), 3.93(t, 2H), 4.2(s, 2H), 6.4(d,1H), 6.87(t, 1H), 7.23(t, 2H), 7.55(d, 1H), 7.73(d, 2H), 8.06(d, 1H),9.07(s, 1H) 340 Meth 20 ¹NMR taken at 100° C. ²Product purified bychromatography eluting with DCM/MeOH (98:2 increasing in polarity to94:6) prior to trituration.Method 11

2-Anilinopyrimidine-4-carbaldehyde oxime

A suspension of 2-anilinopyrimidin-4-carbaldehyde (Method 3; 10 g, 50mmol) and hydroxylamine hydrochloride (17.4 g, 250 mmol) in EtOH (360ml) and pyridine (17.4 g, 215 mmol) was stirred and heated at reflux for1 hour. The reaction mixture was allowed to cool and concentrated toabout 150 ml total volume by evaporation. Distilled water (600 ml) wasadded and the precipitated solid collected by filtration, washedthoroughly with distilled water and dried to give the title compound(9.86 g, 92%) as a white solid. NMR: 6.92 (t, 1H), 7.08 (d, 1H), 7.23(t, 2H), 7.75 (d, 2H), 7.93 (s, 1H), 8.45 (d, 1H), 9.67 (s, 1H), 12.1(s, 1H); m/z 215.

Method 12

2-Anlinoaminomethylpyrimidine

Raney nickel (1.6 g of a 50% suspension in water) was added to asuspension of 2-anilinopyrimidine-4-carbaldehyde oxime (Method 11; 8.8g, 41 mmol) in EtOH (100 ml) and liquid ammonia (10ml). The mixture wasstirred under an atmosphere of hydrogen at 25° C. and 5 bar pressure for16 hours. The catalyst was removed by filtration through diatomaceousearth and the filter pad was washed thoroughly with EtOH and thefiltrate evaporated. The residue was triturated with a mixture ofdiethyl ether and isohexane, the solid product collected by filtration,washed with isohexane and dried to give the title compound (7.11 g, 87%)as a pink solid. NMR: 3.3 (s, 2H), 3.7 (s, 2H), 6.9 (d, 2H), 7.23 (t,2H), 7.78 (m, 2H), 8.4 (d, 1H), 9.45 (s, 1H); m/z 201.

Method 13

4-{[(2-Anilinopyrmidin-4-yl)methyl]amino}-4-oxobutanoic acid

A mixture of 2-anilinoaminomethylpyrimidine (Method 12; 6.6 g, 32.5mmol) and succinic anhydride (3.25 g, 32.5 mmol) in dry THF (130 ml) wasstirred under nitrogen for 20 hours. The solvent was removed byevaporation and the residue stirred and triturated with EtOAc for 30minutes. The solid product was collected filtration, washed with EtOAcand dried to give the title compound (8.75 g, 89.7%) as an off whitesolid NMR: 2.5 (m, 4H), 4.23 (d, 2H), 6.73 (d, 1H), 6.95 (t, 1H), 7.26(t, 2H), 7.77 (d, 2H), 8.38 (d, 1H), 8.45 (t, 1H), 9.52 (s, 1H); m/z301.

Method 14

5-{[(2-Anilinopyrimidin-4-yl)methyl]amino}-5-oxopentanoic acid

By an analogous process to Method 13, 2-anilino-4-aminomethylpyrimidine(Method 12)(4.8 g, 24 mmol) was treated with glutaric anhydride (2.74 g,24 mmol) to give the title compound (3.88 g, 51.7%) as a pale brownsolid. NMR: 1.75 (m, 2H), 2.25 (t, 4H), 4.23 (d, 2H), 6.68 (d, 1H), 6.93(t, 1H), 7.27 (t, 2H), 7.76 (d, 2H), 8.4 (d, 2H), 9.53 (s, 1H): m/z 315.

Method 15

2-Anilino-4-(2,5-dioxopyrrolidin-1-ylmethyl)pyrimidine

1-[3-(Dimethylamino)propyl]3-ethylcarbodiimnide hydrochloride (4.44 g,22 mmol) was added in one portion to a stirred mixture of4-{[(2-anilinopyrimidin-4-yl)methyl]amino}-4-oxobutanoic acid (Method13; 6.0 g, 20mmol) and pentafluorophenol (4.05 g, 22 mmol) in DMF (180ml) under nitrogen. The reaction was stirred at ambient temperatureunder nitrogen for 20 hours and then the reaction mixture wasconcentrated by evaporation. The residue was partitioned between EtOAcand water. The layers were separated and the organic layer washed withwater (2×) and saturated brine, dried (Na₂SO₄) and the volatiles removedby evaporation. The crude pentafluoroester was stirred in dry TBF (218ml) under nitrogen at 0° C. and sodium hydride (528 mg of a 60 %dispersion in oil, 22 mmol) added in one portion. The reaction mixturewas stirred at 0° C. for 15 minutes and then 72 hours at ambienttemperature. Acetic acid (1.32 ml) was added and the reaction mixturewas concentrated by evaporation. The residue was partitioned betweenEtOAc and water and the layers separated. The organic layer was washedwith dilute aqueous sodium hydrogen carbonate solution (2×), water andsaturated brine, dried (Na₂SO₄) and the volatiles removed byevaporation. The crude product was purified by chromatography elutingwith EtOAc/isohexane (60:40 and then 100:0). The purified product wastriturated with isohexane, collected by filtration and dried to give thetitle compound (2.65 g, 47%) as a white solid. NMR: 2.75 (s, 4H), 4.55(s, 2H), 6.75 (d, 1H), 6.93 (t, 1H), 7.25 (t, 2H), 7.64 (d, 2H), 8.37(d, 1H), 9.53 (s, 1H); m/z 283.

Method 16

2-Anilino-4-(2.6-dioxopiperidin-1-ylmethyl)pyrimidine

By an analogous process to method 15,5-{[(2-anilinopyrimidin-4-yl)methyl]amino}-5-oxopentanoic acid (Method14; 3.7 g, 11.78) was converted to give the title compound (2.13 g, 61%)as a pale brown solid. NMR: 1.9 (m, 2H), 2.7 (m, 4H), 4.83 (s, 2H), 6.7(d, 1H), 6.93 (t, 1H), 7.25 (t, 2 H), 7.67 (d, 2 H), 8.33 (d, 1H), 9.5(s, 1H); m/z 297.

Method 17

2-Anilino-4-(5-methylpyrolid-2-on-1-ylmethyl)pyrimidine

Methyl magnesium iodide (6.31 ml of 3M solution in ether, 19.14 mmol)was added over 5 minutes to a stirred solution of2-anilino-4-(2,5-dioxopyrrolidin-1-ylmethyl)pyrimidine (Method 15; 1.8g, 6.38 mmol) in dry THF (96 ml) under nitrogen at −70° C. such that thereaction temperature was maintained below −65° C. The reaction wasstirred 1 hour at −70° C. and then at ambient temperature for 90minutes. Saturated ammonium chloride solution (50 ml) was added and themixture stirred vigorously for 5 minutes. The mixture was thenpartitioned between EtOAc and water and the layers separated. Theorganic layer was washed with saturated brine, dried (Na₂SO₄) and thevolatiles removed by evaporation. The residue was purified by flashchromatography eluting with DCM/MeOH (98:2 and then 96.5:3.5) to giveimpure aminal intermediate (0.75 g, 2.58 mmol) which was treated withtriethylsilane (4.95 ml, 31 mmol) in DCM (5.0 ml) cooled to 0° C.Trifluoroacetic acid (1.03 ml, 13.3 mmol) was added to the solution over2 minutes and the mixture stirred at 0° C. for 15 minutes and then atambient temperature for 18 hours. The reaction mixture was diluted withDCM, stirred vigorously and aqueous sodium hydrogen carbonate solutionadded cautiously until no more effervescence was seen. The layers wereseparated and the organic layer washed with aqueous sodium hydrogencarbonate solution, saturated brine, dried (Na₂SO₄) and the volatilesremoved by evaporation. The crude product was purified by absorption onto a SPE SCX ion exchange column which had been pre-equilibrated withDCM/MeOH (20:80). The column was eluted with DCM/MeOH (20:80) to removeneutrals and then with DCM/2M methanolic ammonia (80:20) to give thetitle compound (754 mg, 50%). NMR: 1.13 (d, 3H), 2.6 (m, 1H), 2.3 (m,3H), 3.7 (q, 1H), 4.13 (d, 1H), 4.55 (d, 1H), 6,68 (d, 1H), 6.93 (t,1H): 7.25 (t, 2H), 7.73 (d, 2H), 8.4 (d, 1H), 9.55 (s, 1H); m/z 283.

Method 18

2-Anilino-4-(6-methylpiperidin-2-on-1-ylmethl)pyrimidine

By an analogous process to Method 17,2-anilino-4-(2,6-dioxopiperidin-1-ylmethyl)pyrimidine (Method 16; 2.21g, 7.47 mmol) was treated with methyl magnesium iodide to prepare theintermediate animal which was taken on without chromatographicpurification to give the final product which was purified, after aqueouswork-up, by chromatography eluting with DCM/MeOH (98:2) to give thetitle compound (375 mg, 17%) as a glassy solid. NMR: 1.2 (d, 3H), 1.65(m, 2H), 1.93 (m, 2H), 2.33 (t, 2H), 3.6 (q, 1H), 4.25 (d, 1H), 4.7 (d,1H), 6.65 (d, 1H), 6.93 (t, 1H), 7.25 (t, 2H), 7.76 (d, 2H), 8.4 (d,1H), 9.53 (s, 1H); m/z 297.

Method 19

2-Anilino-4(2-hydroxethylaminomethyl)pyrimidine

A suspension of 2-anilinopyrimidin-4-carbaldehyde (Method 3; 5.0 g, 25.1mmol) and ethanolamine (1.69 g, 27.6 mmol) in MeOH (63 ml) was stirredunder nitrogen at ambient temperature for 72 hours. The volatiles wereremoved by evaporation and the residue dissolved in EtOH (75 ml) and thesolution stirred under nitrogen. Sodium borohydride (1.04 g, 27.6 mmol)was added rapidly in one portion and the mixture stirred for 20 hours atambient temperature. The volatiles were removed by evaporation and theresidue dissolved in diethyl ether and water and the aqueous layeradjusted to pH 5.5 by careful addition of concentrated hydrochloricacid. The layers were separated and the aqueous layer adjusted to pH11.5 by careful addition of 40% aqueous sodium hydroxide solution. Theaqueous layer was then extracted with DCM (×3) and the organic extractscombined, dried (Na₂SO₄) and the solvent removed by evaporation. Theresidue was triturated with DCM/isohexane, the solid product collectedby filtration, washed with isohexane and dried to give the titlecompound (4.12 g, 75%) as cream solid. NMR: 2.62 (t, 2H), 3.28 (s, 2H),3.47 (t, 2H), 4.47 (s, 1H), 6.9 (m, 2H), 7.25 (t, 2H), 7.78 (d, 2H), 8.4(d, 1H), 9.5 (s, 1H); m/z 245.

Method 20

2-Anilino-4-(3-oxomorpholinomethyl)pyrimidine

A solution of chloroacetyl chloride (268 mg, 2.38 mmol) in dry DCM (5ml) was added dropwise over 10 minutes to a stirred solution of2-anilino-4-(2-hydroxyethylaminomethyl)pyrimidine (Method 19; 582 mg,2.38 mmol) and triethylamine (332 μl, 2.38 mmol) in dry DCM (20 ml)under nitrogen. The reaction mixture was then stirred for 20 hours atambient temperature. The mixture was diluted with DCM and washed withwater (×2), saturated brine, dried (Na₂SO₄) and the volatiles removed byevaporation. The residue was dissolved in dry THF (20 ml), sodiumbromide (227 mg, 2.2 mmol) added and the mixture stirred at 0° C. undernitrogen sodium hydride (88 mg of a 60% dispersion in oil) was added inone portion and the reaction stirred at 0° C. for 10 minutes and then atambient temperature for 72 hours. The volatiles were removed byevaporation and the residue partition between EtOAc and water. Thelayers were separated and the organic layer washed with water, saturatedbrine, dried (Na₂SO₄) and the solvent removed by evaporation. Theresidue was purified by chromatography eluting with EtOAc. The purifiedproduct was triturated with diethyl ether, collected by filtration,washed with diethyl ether and dried to give the title compound (438 mg,70%) as a white solid. NMR: 3.43 (t, 2H), 3.87 (t, 2H), 4.13 (s,2H),4.52 (s, 2H), 6.72 (d, 1H), 6.93 (t, 1H), 7.25 (t 2H), 7.75 (d, 2H),8.42 (d, 1H), 9.55 (s, 1H); m/z 285.

Example 12

The following illustrate representative pharmaceutical dosage formscontaining the compound of formula (I), or a pharmaceutically acceptablesalt or in vivo hydrolysable ester thereof (hereafter compound X), fortherapeutic or prophylactic use in humans:

(a): Tablet I mg/tablet Compound X 100 Lactose Ph.Eur 182.75Croscarmellose sodium 12.0 Maize starch paste (5% w/v paste) 2.25Magnesium stearate 3.0

(b): Tablet II mg/tablet Compound X 50 Lactose Ph.Eur 223.75Croscarmellose sodium 6.0 Maize starch 15.0 Polyvinylpyrrolidone (5% w/vpaste) 2.25 Magnesium stearate 3.0

(c): Tablet III mg/tablet Compound X 1.0 Lactose Ph.Eur 93.25Croscarmellose sodium 4.0 Maize starch paste (5% w/v paste) 0.75Magnesium stearate 1.0

(d): Capsule mg/capsule Compound X 10 Lactose Ph.Eur 488.5 Magnesiumstearate 1.5

(e): Injection I (50 mg/ml) Compound X 5.0% w/v 1 M Sodium hydroxidesolution 15.0% v/v 0.1 M Hydrochloric acid (to adjust pH to 7.6)Polyethylene glycol 400 4.5% w/v Water for injection to 100%

(f): Injection II 10 mg/ml Compound X 1.0% w/v Sodium phosphate BP 3.6%w/v 0.1 M Sodium hydroxide solution 15.0% v/v Water for injection to100%

(g): Injection III (1 mg/ml, buffered to pH6) Compound X  0.1% w/vSodium phosphate BP 2.26% w/v Citric acid 0.38% w/v Polyethylene glycol400  3.5% w/v Water for injection to 100%Note

The above formulations may be obtained by conventional procedures wellknown in the pharmaceutical art. The tablets (a)-(c) may be entericcoated by conventional means, for example to provide a coating ofcellulose acetate phthalate.

1. A compound of formula (I):

wherein: Ring A is carbocyclyl or heterocyclyl fused to the imidazolering; R¹ is halo, nitro, cyano, hydroxy, amino, carboxy, carbamoyl,mercapto, C₁₋₆alkyl, C₁₋₆alkoxy, C₂₋₆alkenyl or C₂₋₆alkynyl; p is 0-4;wherein the values of R¹ may be the same or different; R² is sulphamoylor a group R^(a)—R^(b)—; q is 0-2; wherein the values of R² maybe thesame or different; and wherein p+q=0-5; R³ is halo, nitro, cyano,hydroxy, trifluoromethyl, trifluoromethoxy, amino, carboxy, carbamoyl,mercapto, sulphamoyl, C₁₋₃alkyl, C₂₋₃alkenyl, C₂₋₃alkynyl, C₁₋₃alkoxy,C₁₋₃alkanoyl, N—(C₁₋₃alkyl)amino, N,N—(C₁₋₃alkyl)₂amino,C₁₋₃alkanoylamino, N—(C₁₋₃alkyl)carbamoyl, N,N—(C₁₋₃alkyl)₂carbamoyl,C₁₋₃alkyl S(O)_(a) wherein a is 0 to 2, N—(C₁₋₃alkyl)sulphamoyl orN,N—(C₁₋₃alkyl)₂sulphamoyl; wherein R³ may be optionally substituted oncarbon by one or more R^(c); n is 0 to 2, wherein the values of R³ maybe the same or different; R⁴ and R⁵ are independently selected fromhydrogen, halo, nitro, cyano, hydroxy, trifluoromethoxy, amino, carboxy,carbamoyl, mercapto, sulphamoyl, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₁₋₆alkoxy, C₁₋₆alkanoyl, C₁₋₆alkanoyloxy, N—(C₁₋₆alkyl)amino,N,N—(C₁₋₆alkyl)₂amino, C₁₋₆alkanoylamino, N—(C₁₋₆alkyl)carbamoyl,N,N—(C₁₋₆alkyl)₂carbamoyl, C₁₋₆alkylS(O)_(a) wherein a is 0 to 2,C₁₋₆alkoxycarbonyl, N—(C₁₋₆alkyl)sulphamoyl, N,N—(C₁₋₆alkyl)₂sulphamoyl,C₁₋₆allylsulphonylamino, C₃₋₈cycloalkyl or a 4-7 membered saturatedheterocyclic group; wherein R⁴ and R⁵ may be optionally substituted oncarbon by one or more R^(e); and wherein if said 4-7 membered saturatedheterocyclic group contains an —NH— moiety that nitrogen may beoptionally substituted by a group selected from R^(f); m is 0-4; whereinthe values of R⁴ may be the same or different; R^(a) is selected fromC₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₈cycloalkyl,C₃₋₈cycloalkylC₁₋₆alkyl, phenyl, a heterocyclic group, phenylC₁₋₆alkylor (heterocyclic group)C₁₋₆alkyl; wherein R^(a) may be optionallysubstituted on carbon by one or more R^(g); and wherein if saidheterocyclic group contains an —NH— moiety that nitrogen may beoptionally substituted by a group selected from R^(h); R^(b) is —C(O)—,—N(R^(m))C(O)—, —C(O)N(R^(m))—, —S(O)_(r)—, —OC(O)N(R^(m))SO₂—,—SO₂N(R^(m))— or —N(R^(m))SO₂—; wherein R^(m) is hydrogen or C₁₋₆alkyloptionally substituted by one or more R^(i) and r is 1-2; R^(g) andR^(i) are independently selected from halo, nitro, cyano, hydroxy,amino, carboxy, carbamoyl, mercapto, sulphamoyl, C₁₋₆alkyl, C₂₋₆alkenyl,C₂₋₆alkynyl, C₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkoxy,C₁₋₆alkoxyC₁₋₆alkoxyC₁₋₆alkoxy, C₁₋₆alkanoyl, C₁₋₆alkanoyloxy,N—(C₁₋₆alkyl)amino, N,N—(C₁₋₆alkyl)₂amino, C₁₋₆alkanoylamino,N—(C₁₋₆alkyl)carbamoyl, N,N—(C₁₋₆alkyl)₂carbamoyl, C₁₋₆alkylS(O)_(a)wherein a is 0 to 2, C₁₋₆alkoxycarbonyl, N—(C₁₋₆alkyl)sulphamoyl,N,N—(C₁₋₆alkyl)₂sulphamoyl, C₁₋₆alkylsulphonylamino, C₃₋₈cycloalkyl,phenyl, heterocyclic group, phenylC₁₋₆alkyl-R^(o)—, (heterocyclicgroup)C₁₋₆alkyl-R^(o)—, phenyl-R^(o)— or (heterocyclic group)-R^(o)—;wherein R^(g) and R^(i) independently of each other may be optionallysubstituted on carbon by one or more R^(j); and wherein if saidheterocyclic group contains an —NH— moiety that nitrogen may beoptionally substituted by a group selected from R^(k); R^(o) is —O—,—N(R^(p))—, —C(O)—, —N(R^(p))C(O)—, —C(O)N(R^(p))—, —S(O)_(s)—,—SO₂N(R^(p))— or —N(R^(p))SO₂—; wherein R^(p) is hydrogen or C₁₋₆alkyland s is 0-2; R^(f), R^(h) and R^(k) are independently selected fromC₁₋₄alkyl, C₁₋₄alkanoyl, C₁₋₄alkylsulphonyl, C₁₋₄alkoxycarbonyl,carbamoyl, N—(C₁₋₄alkyl)carbamoyl, N,N—(C₁₋₄alkyl)carbamoyl, benzyl,benzyloxycarbonyl, benzoyl and phenylsulphonyl; wherein R^(f), R^(h) andR^(k) independently of each other may be optionally substituted oncarbon by on or more R^(l); and R^(c), R^(e), R^(l) and R^(j) areindependently selected from halo, nitro, cyano, hydroxy,trifluoromethoxy, trifluoromethyl, amino, carboxy, carbamoyl, mercapto,sulphamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy,methylamino, ethylamino, dimethylamino, diethylamino,N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl,N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl,methylthio, ethylthio, methylsulphinyl, ethylsulphinyl, mesyl,ethylsulphonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulphamoyl,N-ethylsulphamoyl, N,N-dimethylsulphamoyl, N,N-diethylsulphamoyl orN-methyl-N-ethylsulphamoyl; or a pharmaceutically acceptable salt or anin vivo hydrolysable ester thereof.
 2. A compound of formula (I)according to claim 1 wherein Ring A is cyclopentyl, cyclohexyl ormorpholino fused to the imidazole ring or a pharmaceutically acceptablesalt or an in vivo hydrolysable ester thereof.
 3. A compound of formula(I) according to claim 1 wherein R¹ is halo or a pharmaceuticallyacceptable salt or an in vivo hydrolysable ester thereof.
 4. A compoundof formula (I) according to claim 1 wherein p is 0 or a pharmaceuticallyacceptable salt or an in vivo hydrolysable ester thereof.
 5. A compoundof formula (I) according to claim 1 wherein R² is a group R^(a)—R^(b)—;wherein R^(a) is selected from C₁₋₆alkyl optionally substituted oncarbon by one or more R^(g); R^(b) is —N(R^(m))SO₂—; wherein R^(m) ishydrogen; and R^(g) is C₁₋₆alkoxy or a pharmaceutically acceptable saltor an in vivo hydrolysable ester thereof.
 6. A compound of formula (I)according to claim 1 wherein q is 0 or 1 or a pharmaceuticallyacceptable salt or an in vivo hydrolysable ester thereof.
 7. A compoundof formula (I) according to claim 1 wherein R³ is halo or apharmaceutically acceptable salt or an in vivo hydrolysable esterthereof.
 8. A compound of formula (I) according to claim 1 wherein n is0 or a pharmaceutically acceptable salt or an in vivo hydrolysable esterthereof.
 9. A compound of formula (I) according to claim 1 wherein R⁴ ishydrogen or C₁₋₆alkyl or a pharmaceutically acceptable salt or an invivo hydrolysable ester thereof.
 10. A compound of formula (I) accordingto claim 1 wherein m is 0 or 1 or a pharmaceutically acceptable salt oran in vivo hydrolysable ester thereof.
 11. A compound of formula (I)according to claim 1 wherein: Ring A is cyclopentyl, cyclohexyl ormorpholino fused to the imidazole ring; p is 0; R² isN-(2-methoxyethyl)sulphamoyl or N-(2-ethoxyethyl)sulphamoyl; q is 0 or1; n is 0; R⁴ is hydrogen or methyl; and R⁵ is hydrogen; m is 0 or 1; ora pharmaceutically acceptable salt or an in vivo hydrolysable esterthereof.
 12. A process for preparing a compound of formula (I),according to claim 1, or a pharmaceutically acceptable salt or an invivo hydrolysable ester thereof which process (wherein variable groupsare, unless otherwise specified, as defined in claim 1) comprises:Process a) reacting a pyrimidine of formula (II):

wherein L is a displaceable group; with an aniline of formula (III):

or Process b) reacting a compound of formula (IV):

with a compound of formula (V):

wherein T is O or S; R^(x) may be the same or different and is selectedfrom C₁₋₆alkyl; or Process c) for compounds of formula (I) where R² issulphamoyl or a group R^(a)—R^(b)— and R^(b) is —NHSO₂—; reacting apyrimidine of formula (VI):

wherein X is a displaceable group; with ammonia or an amine of formula(VII):R^(a)—NH₂   (VII); or Process d) for compounds of formula (I); reactinga pyrimidine of formula (VIII)

with a compound of formula (IX):

where Y is a displaceable group; or Process e) cyclizing a compound offormula (X):

and thereafter optionally: i) converting a compound of the formula (I)into another compound of the formula (I); ii) removing any protectinggroups; iii) forming a pharmaceutically acceptable salt or in vivohydrolysable ester.
 13. A pharmaceutical composition which comprises acompound of the formula (I), or a pharmaceutically acceptable salt or invivo hydrolysable ester thereof, according to claim 1, in associationwith a pharmaceutically-acceptable diluent or carrier.
 14. A method oftreating rheumatoid arthritis, in a warm-blooded animal in need of suchtreatment, which comprises administering to said animal an effectiveamount of a compound of formula (I) or a pharmaceutically acceptablesalt or in vivo ester thereof, as claimed in claim 1.